The vast majority of patients with pancreatic cancer are not cured by resection alone. Standard therapies are associated with low response rates and modest improvements in survival, and there is a dire need for more effective therapies. Aptamers are a class of therapeutic nucleic acid (RNA or DNA) molecules, which specifically bind to existing target proteins. Aptamers are generated by an iterative screening process of large combinatorial libraries that can be modified for nuclease resistance. Aptamers can have direct therapeutic effects mediated by binding their protein targets. In addition, aptamers that bind to cell surface receptors can be internalized by cells expressing those receptors and be utilized to deliver other therapeutic cargo. Our global hypothesis is that aptamers that bind targets over-expressed on pancreatic cancer cells relative to normal cells can be used to selectively deliver cytotoxic cargo, such as small interfering RNAs (siRNAs) or chemotherapeutic agents, to pancreatic cancer cells. We have 2 specific aims: Specific Aim #1: To utilize selection strategies against complex targets to identify new pancreatic cancer targets. The ideal target for aptamer-mediated delivery is one that is highly expressed on the surface of all pancreatic cancers, efficiently internalized, and not expressed on the surface of normal cells. Selection strategies utilizing complex targets such as whole cancer cells or tumor tissue in vivo allow the aptamers to choose their own targets. This aim includes in vitro selection against whole pancreatic cancer cells as well as in vivo selection against pancreatic cancer xenografts and genetically engineered mouse models of pancreatic cancer. We will characterize selected aptamers by identifying their specific protein and cellular targets. This aim therefore has the potential to simultaneously identify novel targets and the agents to mediate delivery to them. Aptamers that are internalized by pancreatic cancer cells will be further evaluated in Aim #2. Specific Aim #2: To utilize aptamers that is internalized by pancreatic cancer cells for specific delivery of siRNAs and other therapeutic cargo. We have preliminary data demonstrating that a RNA aptamer that binds EGFR and a DNA aptamer that binds nucleolin are internalized by pancreatic cancer cells. In parallel with the identification of new aptamers in Aim #1, we will utilize these extant aptamers to optimize constructs for delivery of K-ras siRNAs and gemcitabine polymers into cells. Then, we will use aptamers discovered in Aim #1 with the cargo technology gained in the first part of Aim #2 to formulate more specific and effective constructs to deliver K-ras siRNAs and gemcitabine polymers into pancreatic cancer cells in vitro and in vivo. PUBLIC HEALTH RELEVANCE: Standard therapies for pancreatic cancer (surgical resection, chemotherapy, and radiation) are not very effective. The objective of this project is to develop novel agents that target proteins that are present at higher levels on pancreatic cancer cells than on normal cells and that can be used to selectively deliver toxic cargo to pancreatic cancer cells. Such agents would be used in combination with standard therapies to improve survival in patients with advanced disease and cure a larger percentage of patients with respectable disease.